In vitro cytotoxic and genotoxic effects of Cissus verticillata and Sphagneticola trilobata used for treatment of Diabetes Mellitus in Brazilian folk medicine

Cissus verticillata and Sphagneticola trilobata have been used in Brazilian folk medicine for Diabetes Mellitus treatment, although their pharmacological and toxicological profile has not been clearly established. Thus, the aim of this study was to evaluate the preclinical toxicity of the aqueous extracts of C. verticillata and S. trilobata. The main groups of secondary metabolites were investigated, and the species differed by the presence of coumarins in C. verticillata and by tannins in S. trilobata extracts. The highest contents of phenolic compounds and flavonoids were quantified in C. verticillata infusion with 2.594 ± 0.04 mg equivalents of gallic acid g-1 of extract and 1.301 ± 0.015 mg equivalents of catechin g-1 of extract, respectively. While the extract of S. trilobata showed minimum values of these compounds, with 0.002 ± 0.001 mg equivalents of gallic acid g-1 extract and 0.005 ± 0.0004 mg equivalents of catechin g-1 of extract, respectively. These differences implied the results of in vitro antioxidant activity evaluated using ferric reducing antioxidant power (FRAP), in which the sample of C. verticillata at 5 mg mL-1 showed a value of 122 µM ferrous sulfate equivalents (FSE), while S. trilobata showed 0.93 µM FSE at the same concentration. With respect to cytotoxic assay with murine fibroblast cell line (3T3) only S. trilobata exhibited cytotoxic effects measured by MTT and Sulforhodamine B assays, evidenced by the cell viability value of approximately 16%, in both tests after 24 and 72 hours of exposure of the cells to 5 mg mL-1 of the extract. Comparatively, at 5 mg mL-1 the C. verticillata extract showed cell viability of 142% and 95%, respectively, after 24 hours of cell exposure. On the other hand, both species showed genotoxic profiles evidenced by chromosomal aberrations by Allium cepa bioassay, observed by the higher percentage values of chromosome bridges, chromosome loss, and disturbed anaphase for all concentrations of both extracts than those of the negative control. The results support the characterization of the toxicological profile for both species and create an alert regarding the use of S. trilobata, which should be avoided.

Estudo toxicológico pré-clínico agudo com o extrato hidroalcoólico das folhas de Cissus sicyoides L. (Vitaceae) / Acute pre-clinical toxicological study with the hydroalcoholic extract of the leaves of Cissus sicyoides L. (Vitaceae)

O estudo objetivou realizar ensaio toxicológico pré-clínico inicial para investigar a toxicidade da planta. Na investigação da DL50 foram utilizados grupos de camundongos Swiss machos (n = 10) submetidos a doses de 5,0 g/kg, via oral, e dose de 2,0 g/kg por via intraperitoneal, da fração aquosa das folhas (FAF) de Cissus sicyoides L. Foi retirado sangue através do plexo braquial após sete dias, para análise laboratorial de parâmetros hematológicos e bioquímicos. No estudo anatomopatológico, os animais foram examinados macro e microscopicamente, com ressecção de pulmão, coração, fígado e rins. Para análise dos dados foram utilizadas técnicas de estatística descritiva e inferencial. Na avaliação dos parâmetros hematológicos e bioquímicos dos camundongos Swiss, foram detectadas alterações nas transaminases AST e ALT e na fosfatase alcalina (FAL), caracterizando as alterações hepáticas demonstradas no estudo histopatológico. Na avaliação histológica do fígado de camundongos, evidenciou-se hepatite reacional com portite linfocitária crônica e lobular multifocal, hiperplasia kupferiana, colapsos focais da trama reticular, ausência de fibrose portal e lobular. O estudo toxicológico pré-clínico agudo demonstrou em camundongos alterações hepáticas.

The aim of the study was to carry through pre-clinical toxicological assay to analyze the toxicity of the plant. In the inquiry of the LD50 groups of male Swiss rats (n = 10) were given orally 5.0 g/kg, and 2.0 g/kg intraperitoneally, of the water fraction of the leaves (WFL) of Cissus sicyoides L. Blood was removed through brachial plexus after seven days, for laboratory analysis of hematological and biochemical parameters. In the anatomopathological study, the animals had been examined macro and microscopically, with dissection of lung, heart, liver and kidneys. For analysis of the data techniques of descriptive and inferencial statistics had been used. In the evaluation of the hematological and biochemical parameters of the Swiss rats, alterations in aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (FAL) had been detected, characterizing the demonstrated hepatic alterations in the histopathological study. In the histological evaluation of the rats's liver, submitted to the poisoning, reactionary hepatitis with chronic linfocity portite was proven and to lobular multifocal, hiperplasia kupferiana, focal collapses of the reticular tram, absence of fibrosis, absences of cellular atipias and reactionary subcapsulite. The acute pre-clinical toxicological study demonstrated hepatic alterations in rats.